Abstract Many cancer types including pancreatic cancer have very high mortality rates because it is difficult to detect their presence in the early stage. Current biomarkers for pancreatic cancer such as serum CA-19-9 protein face high false positive and negative rates. The proposed studies seek to develop a robust and user-friendly novel molecular beacon and lipoplex nanoparticle biochip technology and validate its clinic potential by detecting Glypican 1 mRNA and protein in extracellular vesicles from pancreatic cancer patient serum samples for imminent use in the field of early cancer diagnosis. The specific aims of this project are: Aim 1: Development of Robust Biochip Technology. 1-1. Improved MBs for high stability and mis-match specificity; 1-2. Standard vesicles (SVs) for chip and analyzer calibration and analytical quantification; 1-3. Non-TIRF fluorescence microscope as a low-cost analyzer and automated software module for fast and user-friendly image acquisition and analysis. Specific Aim 2: Multi-site Technology Validation for Pancreatic Cancer Diagnosis. 2-1. Comparison with qRT-PCR, NGS and ELISA for performance, cost, assay time, and user friendliness. 2-2. Multi-site blind validation: Lab-to-lab repeatability and blind validation comparison at OSU and MSKCC using patient and normal donor serum samples.